HASHIMOTO Tamotsu (GOTOH) Kyoto Prefectural University of Medicine, medical department, Associate Professo, 医学部, 助教授 (00237942)
NAKAJIMA Kenji Kyoto Prefectural University of Medicine, medical department, Professor, 医学部, 教授 (00237265)
|Budget Amount *help
¥2,100,000 (Direct Cost : ¥2,100,000)
Fiscal Year 1997 : ¥400,000 (Direct Cost : ¥400,000)
Fiscal Year 1996 : ¥400,000 (Direct Cost : ¥400,000)
Fiscal Year 1995 : ¥1,300,000 (Direct Cost : ¥1,300,000)
The PCR-RFLP method is widely used for detecting genotypes of Apolipoprotein E (ApoE). However, there are some problems to detect the heterozygous pattern of ApoE by PCR-RFLP method. When the most popularly used PCR primers were employed, the density of restriction fragments vary depending on genotypes of the gene. Also, there are some problems to detect heterozygous pattern of ApoE by direct sequencing with PCR.We prepared new PCR primers for detecting genotypes of ApoE.After amplification by PCR,genotypes of Apolipoprotein were determined by RFLP method and direct sequencing. When the new primers were used, density of fragment after digestion by restriction enzyme was identical and heterozygous pattern was easily identified by direct sequencing. These results indicated that the new primers were more reliable for detecting genotypes of ApoE than ordinal primers.
We performed the genotype analysis of ApoE in the northern part of Kyoto Prefecture using these new primers for ApoE.Subjects were 198 residents 65 years old and older in the town, 21 eldely in a hospital. These 219 persons were divided into three groups ; no dementia group 183, senile dementia of Alzheimer type (SDAT) group 22 and vascular dementia group 14. The frequencies of ApoE2/2,3/3,2/4,3/4,4/4, were 1.1%, 5.5%, 75.4%, 1.1%, 16.4%, 0.5% in no dementia group and 0%, 0%, 77.2%, 4.5%, 18.2%, 0% in SDAT group. The frequencies of epsilon 4 were 9.3% in no dementia group and 11.4% in SDAT group. The frequency of epsilon 4 was higher in SDAT group than no dementia group, but this difference was not statistically significant. The frequency epsilon 4 in no dementia group in this area was higher reported so far from other Japanese areas, but this difference was not significant, either.